Methods of use of quinolone compounds against anaerobic pathogenic bacteria

ABSTRACT

This invention relates, in part, to newly identified methods of using quinolone antibiotics, particularly a gemifloxacin compound against pathogenic bacteria, especially anaerobic pathogens.

[0001] This invention relates, in part, to newly identified methods ofusing quinolone antibiotics, particularly a gemifloxacin compoundagainst anaerobic bacteria, especially unusual anaerobic bacteria.

BACKGROUND OF THE INVENTION

[0002] Quinolones have been shown to be effective to varying degreesagainst a range of certain anaerobic pathogens. However, as diseasescaused by these pathogens are on the rise, there exists a need forantimicrobial compounds that are more potent than the present group ofquinolones.

[0003] Gemifloxacin mesylate (SB-265805) is a novel fluoroquinoloneuseful as a potent antibacterial agent. Gemifloxacin compounds aredescribed in detail in patent application PCT/KR98/00051 published as WO98/42705. Patent application EP 688772 discloses novelquinoline(naphthyridine)carboxylic acid derivatives, including anhydrous(R,S)-7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylicacid of formula I.

[0004] PCT/KR98100051 discloses(R,S)-7-(3-aminomethyl-4-syn-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylicacid methanesulfonate and hydrates thereof including the sesquihydrate.

[0005] While in vitro testing of new antimicrobial compounds is oftenextensive, these studies tend to focus on a limited range of typicalanaerobic bacterial pathogens (Cormicon, et al., Antimicrob. AgentsChemother., 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect.,4:280-284, 1998; Marco, et al., J. Antimicrob. Chemother., 40:605-607,1997). Moreover, it has been reported that (Goldstein, et al.,Antimicrob. Agents Chemother., Submitted) the activity of gemifloxacinagainst typical anaerobic bacteria; it showed activity againstBacteroides fragilis and certain Prevotella and Porphyromonas strains,but only limited activity against B. thetaiotaomicron, B. distasonis andB. ovatus. Data is lacking about the activities of new quinolonecompounds against many of the less frequently encountered anaerobicpathogens, such as Actinomyces spp., Anaerobiospirrilum spp.,Porphyromonas spp., and Bilophila wadsworthia.

[0006] Provided herein is an invention based, in part, on a significantdiscovery made using a gemifloxacin compound against various unusual,though medically impotant, anaerobic bacteria, demonstrating theactivity of the gemifloxacin compound used was superior to a number ofquinolones as described in more detail herein. Gemifloxacin compoundsare valuable compounds for the treatment of bacterial caused by a rangeof anaerobic pathogens, including those resistant to usual oral therapy,thereby filling an unmet medical need.

SUMMARY OF THE INVENTION

[0007] An object of the invention is a method for modulating metabolismof anaerobic pathogenic bacteria comprising the step of contactinganaerobic pathogenic bacteria with an antibacterially effective amountof a composition comprising a quinolone, particularly a gemifloxacincompound, or an antibacterially effective derivative thereof.

[0008] A further object of the invention is a method wherein saidanaerobic pathogenic bacteria is selected from the group consisting of:a member of the genus Peptostreptococci, a Peptostreptococciasaccharolyticus, a Peptostreptococci magnus, a Peptostreptococcimicros, a Peptostreptococci prevotii, a Porphyromonas asaccharolytica, aPorphyromonas canoris, a Porphyromonas gingivalis, a Porphyromonasmacaccae, a member of the genus Actinomyces, an Actinomyces israelii, anActinomyces odontolyticus, a member of the genus Clostridium, aClostridium innocuum, a Clostridium clostridioforme, a Clostridiumdifficile, a member of the genus Anaerobiospirillum, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a Prevotella intermedia, a Prevotella heparinolytica, aPrevotella oris-buccae, a Prevotella bivia, a Prevotella melaninogenica,a member of the genus Fusobacterium, a Fusobacterium naviforme, aFusobacterium necrophorum, a Fusobacterium varium, a Fusobacteriumulcerans, a Fusobacterium russii, a member of the genus Bilophila, aBilophila wadsworthia.

[0009] Also provided by the invention is a method of treating orpreventing a bacterial infection by anaerobic pathogenic bacteriacomprising the step of administering an antibacterially effective amountof a composition comprising a quinolone, particularly a gemifloxacincompound to a mammal suspected of having or being at risk of having aninfection with anaerobic pathogenic bacteria.

[0010] A preferred method is provided wherein said modulating metabolismis inhibiting growth of said bacteria or killing said bacteria.

[0011] A further preferred method is provided wherein said contactingsaid bacteria comprises the further step of introducing said compositioninto a mammal, particularly a human.

[0012] Further preferred methods are provided by the invention whereinsaid bacteria is selected from the group consisting of: a member of thegenus Peptostreptococci, a Peptostreptococci asaccharolyticus, aPeptostreptococci magnus, a Peptostreptococci micros, aPeptostreptococci prevotii, a member of the genus Porphyromonas, aPorphyromonas asaccharolytica, a Porphyromonas canons, a Porphyromonasgingivalis, a Porphyromonas macaccae, a member of the genus Actinomyces,an Actinomyces israelii, an Actinomyces odontolyticus, a member of thegenus Clostridium, a Clostridium innocuum, a Clostridiumclostridioforme, a Clostridium difficile, a member of the genusAnaerobiospirillum, a member of the genus Bacteroides, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a member of the genus Prevotella, a Prevotella intermedia, aPrevotella heparinolytica, a Prevotella oris-buccae, a Prevotella bivia,a Prevotella melaninogenica, a member of the genus Fusobacterium, aFusobacterium naviforme, a Fusobacterium necrophorum, a Fusobacteriumvarium, a Fusobacterium ulcerans, a Fusobacterium russii, a member ofthe genus Bilophila, a Bilophila wadsworthia.

[0013] Various changes and modifications within the spirit and scope ofthe disclosed invention will become readily apparent to those skilled inthe art from reading the following descriptions and from reading theother parts of the present disclosure.

DESCRIPTION OF THE INVENTION

[0014] The present invention provides, among other things, methods forusing a composition comprising a quinolone, particularly a gemifloxacincompound against anaerobic bacteria.

[0015] As used herein “gemifloxacin compound(s)” means a compound havingantibacterial activity described in patent application PCT/KR98/00051published as WO 98/42705, or patent application EP 688772.

[0016] This invention was based, in part, on analyses evaluating thecomparative activity of gemifloxacin against various anaerobicpathogens. Since anaerobic susceptibility testing is not routinelyperformed in most clinical laboratories, if it is performed at all, theclinician must rely on published studies to help guide both empiricaltherapy as well as specific therapy in situations that involve lesscommonly isolated or identified anaerobes or mixed infections at othersites. Most in vitro studies of gemifloxacin against anaerobic bacteriafocus their attention on common intra-abdominal pathogens such asBacteroides fragilis and Clostridium perfringens or lump the anaerobesinto large groups without speciation (Cormicon, et al., Antimicrob.Agents Chemother., 41:204-211, 1997; Marco, et al., J. Antimicrob.Chemother., 40:605-607, 1997). Consequently there is unmet medical needfor data regarding certain species often recovered from respiratory andgynecological infections as well as less frequently identifiedcomponents of mixed abdominal infections, as well as methods used forther treatment of such pathogens.

[0017] The present invention was based, in part, on analyses showingthat gemifloxacin compared favorably with trovafloxacin against theGram-positive anaerobes tested as well as the other unusual isolatesstudied [see, for example, Table 1]. Cormicon and Jones (Cormicon, etal., Antimicrob. Agents Chemother., 41:204-211, 1997) studied 10 strainsof peptostreptococci and found an MIC₉₀ of 2 ug/ml for gemifloxacinwhich is in contrast to our study which included 45 strains ofpeptostreptococci from 4 species, all of which were susceptible to

0.25 ug/ml of gemifloxacin. The reason for this discrepancy can not beaccounted for by methodological variations since both studies usedbrucella agar and an agar dilution method. Marco, et al. (Marco et al.J. Antimicrob. Chemother., 40:605-607, 1997) studied 18 strains ofpeptostreptococci and also found an MIC₉₀ of 2 ug/ml [range, z,9000.25-8 ug/ml] for gemifloxacin.

[0018] Differences in the susceptibility of different Clostridiumspecies to gemifloxacin were apparent in assays described herein, withC. clostridioforme and C. innocuum being relatively susceptible while C.difficile was often resistant to gemifloxacin. In the current study, theten C. ramosum isolates studied had an MIC₉₀ of 1 ug/ml while a priorstudy (Goldstein, et al., Antimicrob. Agents Chemother, Submitted) theMIC₉₀ for the 14 isolates studied was 8 ug/ml. With the exception of twostrains, all isolates in these two studies were different and most camefrom blood cultures. The apparent disparity comes from the higher MICsof {fraction (3/14)} strains in the prior study and highlights theproblem of testing small numbers of isolates of a single species, whichhas been solved by the analyses underpinning certain embodiments of thecurrent invention. Further, Cormicon and Jones (Cormicon, et al.,Antimicrob. Agents Chemother., 41:204-211, 1997) studied ten clostridialisolates and found a maximum MIC of 2 ug/ml to gemifloxacin. Marco, etal. (Marco, et al., J. Antimicrob. Chemother. 40:605-607, 1997) reportedall 19 unspecieted clostridial isolates they studied to be susceptibleto

2 ug/ml.

[0019] The data presented herein shows that there is marked variation insusceptibility patterns of different anaerobic genera and species totrovafloxacin and gemifloxacin and that important clinical anaerobicisolates should have individual strain susceptibilities determined. Itis difficult to predict susceptibility based on a grouping of severalspecies in a less commonly encountered or identified genus, and thisproblem has been solved in the methods of the invention.

[0020] Gemifloxacin exhibited good activity against Gram-positiveanaerobes, especially the four Peptostreptococcus species tested, aswell as the Porphyromonas species tested.

[0021] As provided herein, activities of gemifloxacin and comparatorantimicrobial agents were determined by an agar dilution method against419 clinical strains of less commonly identified, though medicallyimportant, species of anaerobes. Gemifloxacin was generally more activethan trovafloxacin against Gram-positive strains by one to twodilutions. Peptostreptococci [P. asaccharolyticus, P. magnus, P. micros,and P. prevotii] and Porphyromonas spp. [P. asaccharolytica, P. canoris,P. gingivalis, P. macaccae] were all susceptible to

0.25 ug/ml of gemifloxacin. Actinomyces israelii, Actinomycesodontolyticus, Clostridium innocuum, Clostridium clostridioforme,Anaerobiospirillum spp., Bacteroides tectum, B. ureolyticus, B. gracilis[now Campylobacter gracilis], Prevotella intermedia, Prevotellaheparinolytica, Prevotella oris-buccae group had MIC₉₀s of

2

g/ml. Fusobacterium naviforme and F. necrophorum were also susceptibleto

2

g/ml, while F. varium strains exhibited a bimodal pattern; the otherFusobacterium species, such as F. ulcerans, F. russii, as well asVeillonella spp., Prevotella melaninogenica group, P. bivia, Clostridiumdifficile, and Bilophila wadsworthia were relatively resistant togemifloxacin [MIC₉₀s

4 ug/ml].(See Table 1). TABLE 1 In vitro activity [μg/ml] gemifloxacin,trovafloxacin, and other oral antimicrobial agents against unusualanaerobic pathogens. Minimal Inhibitory Concentration Organism & Agent(no. isolates)^(A) Range 50% 90% Actinomyces odontolyticus [10]Gemifloxacin 1-2 2 2 Trovafloxacin 2-4 4 4 Penicillin G 0.125-0.25 0.125.0125 Amoxicillin clavulanate 0.06-0.125 0.125 0.25 Clindamycin

0.015-0.5 0.125 0.25 Erythromycin

0.015-0.03

0.015 0.03 Azithromycin

0.015-0.06 0.03 0.06 Clarithromycin

0.015

0.015

0.015 Metronidazole 4->32 16 32 Actinomyces israelii [6] Gemifloxacin0.5-2 1 Trovafloxacin 0.5-2 1 Penicillin G

0.015-0.25 0.03 Amoxicillin clavulanate 0.03-1 0.03 Clindamycin0.06-0.25 0.06 Erythromycin 0.03 0.03 Azithromycin 0.06 0.06Clarithromycin

0.015

0.015 Metronidazole 1-32 4 Anaerobiospirillum thomasii [13] Gemifloxacin0.06-0.25 0.125 0.125 Trovafloxacin 0.06-0.5 0.125 0.25 Penicillin G0.06-0.125 0.06 0.125 Amoxicillin clavulanate 0.125-0.25 0.125 0.25Clindamycin 8->32 32 >32 Erythromycin 1-16 4 8 Azithromycin 0.125-1 0.51 Clarithromycin 2-32 4 16 Metronidazole 1-4 2 4 Anaerobiospirillumsucciniciproducens [3] Gemifloxacin 0.5-2 1 Trovafloxacin 0.5-2 1Penicillin G 0.5-1 0.5 Amoxicillin clavulanate 0.25-0.5 0.25 Clindamycin32 32 Erythromycin 8-16 16 Azithromycin 0.5-1 0.5 Clarithromycin 8-32 32Metronidazole 4-8 8 Bacteroides gracilis [11] Gemifloxacin

0.015-1

0.015 1 Trovofloxacin

0.015-2 0.03 0.5 Penicillin G

0.015-4 0.125 4 Amoxicillin clavulanate 0

.015-2 0.5 2 Clindamycin 0.03-8 0.25 2 Erythromycin 0.125-2 1 2Azithromycin 0.06-0.5 0.125 0.5 Clarithromycin 0.25-2 1 1 Metronidazole0.06 > 32 0.5 >32 Bacteroides tectum [22] Gemifloxacin 0.06-8 0.125 0.25Trovafloxacin 0.03-0.125 0.06 0.125 Penicillin G

0.015-32 0.03 16 Amoxicillin clavulanate 0.03-0.5 0.06 0.5 Clindamycin

0.015-0.125

0.015

0.015 Erythromycin 0.25-1 0.5 0.5 Azithromycin 0.5-2 1 2 Clarithromycin0.125 0.125 0.125 Metronidazole 0.125-2 0.5 0.5 Bacteroides ureolyticus[17] Gemifloxacin

0.015-2

0.015 2 Trovafloxacin

0.015-4 0.06 4 Penicillin G

0.015-1

0.015 0.25 Amoxicillin clavulanate

0.015-1

0.015 0.125 Clindamycin 0.03-0.5 0.06 0.25 Erythromycin 0.125-2 .025 2Azithromycin 0.06-0.25 0.06 0.25 Clarithromycin 0.125-4 0.5 2Metronidazole 0.06-2 0.25 1 Bilophila wadsworthia [16] Gemifloxacin0.125->8 0.25 4 Trovofloxacin 0.125->8 0.5 >8 Penicillin G 2-16 4 8Amoxicillin clavulanate 1-4 2 4 Clindamycin 0.25-2 0.5 2 Erythromycin4-32 16 32 Azithromycin 1-16 4 16 Clarithromycin 4-32 16 32Metronidazole 0.125 0.125 0.125 Clostridium clostridioforme [11]Gemifloxacin 0.5->8 0.5 1 Trovafloxacin 1-8 4 4 Penicillin G 0.5->32 116 Amoxicillin clavulanate 0.5-8 0.5 1 Clindamycin

0.015-2 0.06 2 Erythromycin 0.25->32 16 >32 Azithromycin 0.125->3216 >32 Clarithromycin 0.125->32 4 >32 Metronidazole 0.03-1 0.125 0.5Clostridium difficile [14] Gemifloxacin 1->8 2 >8 Trovafloxacin 0.5->81 >8 Penicillin G 1-4 2 4 Amoxicillin clavulanate 0.5-1 1 1 Clindamycin0.25->32 0.5 >32 Erythromycin 0.25 > 32 0.5 >32 Azithromycin 1->32 2 >32Clarithromycin 0.125->32 0.5 >32 Metronidazole 0.25-1 0.5 0.5Clostridium inocuum [11] Gemifloxacin 0.125->8 0.25 2 Trovafloxacin0.25->8 0.5 8 Penicillin G 0.25->3 0.5 0.5 Amoxicillin clavulanate 0.5-20.5 0.5 Clindamycin 0.25->32 0.5 >32 Erythromyccin 0.5->32 >32 >32Azithromycin 0.125->32 >32 >32 Clarithromycin 0.25->32 >32 >32Metronidazole 0.5-2 0.5 1 Clostridium ramosum [10] Gemifloxacin 0.125-20.25 1 Trovafloxcin 0.25-8 0.5 2 Pencillin G 0.06-1 0.06 1 Amoxicillinclavulanate 0.06-0.25 0.06 0.25 Clindamycin 0.25-4 2 2 Erythromycin0.5->32 1 >32 Azithromycin 0.125->32 0.25 >32 Clarithromycin 0.25->320.5 >32 Metronidazole 1 1 1 Fusobacterium spp group 1 [19]^(B)Gemifloxacin 0.06-8 0.25 8 Trovafloxacin 0.25-4 0.5 4 Penicillin G

0.015-16

0.015 2 Amoxicillin clavulanate

0.015-0.25 0.06 0.125 Clindamycin

0.015-2 0.06 0.125 Erythromycin 1->32 8 32 Azithromycin 0.06-32 1 8Clarithromycin

0.015-32 8 32 Metronidazole 0.125-0.5 0.25 4 Fusobacterium spp. group 2[12]^(C) Gemifloxacin 0.125->8 4 4 Trovafloxacin 1->8 4 4 Penicillin G

0.015->32 0.25 0.5 Amoxicillin clavulanate 0.125->4 1 2 Clindamycin0.06-8 1 8 Erythromycin 8->32 >32 >32 Azithromycin 1->32 16 32Clarithromycin 4->32 >32 >32 Metronidazole 0.125-1 0.5 1 Fusobacteriumrussii Gemifloxacin 0.5->8 >8 >8 Trovafloxacin 0.5-4 4 4 Penicillin G

0.015-0.06 0.03 0.06 Amoxicillin clavulanate

0.015-0.25 0.06 0.125 Clindamycin

0.015-0.125 0.03 0.06 Erythromycin 1->32 4 >32 Azithromycin 0.03-32 0.2532 Clarithromycin 2->32 4 >32 Metronidazole

0.015-0.25 0.125 0.25 Fusobacterium varium [17] Gemifloxacin0.25->8 >8 >8 Trovafloxacin 0.5->8 4 >8 Penicillin G 0.03->32 0.5 8Amoxicillin clavulanate 0.125-4 2 4 Clindamycin 0.06-16 4 16Erythromycin 32->32 >32 >32 Azithromycin 2->32 32 >32 Clarithromycin32->32 >32 >32 Metronidazole 0.125-4 1 2 Peptostreptococcusasaccharolyticus [11] Gemifloxacin 0.125-0.25 0.25 0.25 Trovafloxacin0.5-2 1 1 Penicillin G

0.015-1 0.03 0.25 Amoxicillin clavulanate 0.03-1 0.03 0.125 Clindamycin

0.015->32 0.06 >32 Erythromycin 1->32 4 >32 Azithromycin 0.5->32 4 >32Clarithromycin 0.5->32 2 >32 Metronidazole 0.125-2 0.5 1Peptostreptococcus magnus [13] Gemifloxacin 0.030.03 0.03 0.06Trovafloxacin 0.06-0.25 0.125 0.25 Penicillin G

0.015-1 0.03 0.25 Amoxicillin clavulanate 0.03-1 0.03 0.125 Clindamycin0.06-2 0.5 2 Erythromycin 1->32 4 >32 Azithromycin 2->32 4 >32Clarithromycin 0.5->32 2 >32 Metronidazole 0.25-2 0.5 0.5Peptostreptococcus micros [12] 0.06-0.125 0.06 0.06 GemifloxacinTrovafloxacin 0.03-0.125 0.06 0.06 Penicillin G

0.015-0.03

0.015 0.03 Amoxicillin clavulanate 0.03-0.125 0.03 0.125 Erythromycin0.5-1 0.5 0.5 Azithromycin 0.5-1 0.5 1 Clarithromycin 0.6 0.5 0.5Clindamycin 0.06-0.125 0.125 0.125 Metronidazole 0.03-0.25 0.25 0.25Peptostreptococcus prevotii [9] Gemifloxacin 0.06-0.25 0.125 —Trovafloxacin 0.25-1 0.25 — Penicillin G 0.03-0.06 0.03 — Amoxicillinclavulanate

0.015-0.125 0.03 — Clindamycin 0.030-32 1 — Erythromycin 0.03->32 >32 —Azithromycin 0.06->32 32 — Clarithromycin

0.015->32 >32 — Metronidazole 0.125-1 0.5 — Porphyromonasasaccharolyticus [11] Gemifloxacin 0.06-0.125 0.06 0.125 Trovafloxacin0.03-0.25 0.25 0.25 Penicillin G

0.015

0.015

0.015 Amoxicillin clavulanate

0.015-0.03

0.015 0.03 Clindamycin

0.015->32

0.015 >32 Erythromycin 0.03-32 0.03 32 Azithromycin 0.125->32 0.25 >32Clarithromycin

0.015->32 0.06 >32 Metronidazole

0.015

0.015

0.015 Porphyromonas canoris [10] Gemifloxacin 0.06-0.25 0.25 0.25Trovafloxacin 0.06-0.5 0.25 0.5 Penicillin G

0.015-0.03

0.015

0.015 Amoxicillin clavulanate

0.015-0.03

0.015 0.03 Clindamycin

0.015

0.015

0.015 Erythromycin 0.03-0.25 0.06 0.125 Azithromycin 0.125-0.5 0.25 0.25Clarithromycin 0.06-0.125 0.06 0.125 Metronidazole

0.015-0.5 0.25 0.25 Porphyromonas gingivalis [13] Gemifloxacin

0.015-0.125 0.06 0.125 Trovafloxacin 0.03-0.06 0.06 0.06 Penicillin G

0.015-0.06

0.015 0.03 Amoxicillin clavulanate

0.015-0.06

0.015 0.06 Clindamycin

0.015

0.015

0.015 Erythromycin 0.06-0.5 0.125 0.5 Azithromycin 0.125-1 0.25 0.5Clarithromycin 0.06-0.125 0.06 0.125 Metronidazole

0.015-0.03

0.015 0.03 Porphyromonas macaccae [13] Gemifloxacin 0.03-0.125 0.060.125 Trovafloxacin 0.03-0.125 0.06 0.125 Penicillin G

0.015-1 0.5 0.5 Amoxicillin clavulanate

0.015-0.06

0.015

0.015 Clindamycin

0.015-0.03

0.015

0.015 Erythromycin 0.06-0.25 0.125 0.25 Azithromycin 0.125-1 0.5 0.5Clarithromycin 0.06-0.125 0.125 0.125 Metronidazole

0.015-0.125 0.06 0.125 Porphyromonas spp. [11]^(D) Gemifloxacin0.06-0.125 0.06 0.125 Trovafloxacin 0.06-1 0.25 1 Penicillin G

0.015-4

0.015

0.015 Amoxicillin clavulanate

0.015-0.06

0.015

0.015 Clindamycin

0.015

0.015

0.015 Erythromycin

0.015-0.5 0.06 0.06 Azithromycin 0.125-1 0.25 0.5 Clarithromycin0.06-0.125 0.06 0.125 Metronidazole

0.015-0.25 0.03 0.125 Prevotella bivia [21] Gemifloxacin 4->8 8 8Trovafloxacin 1-4 2 2 Penicillin G 0.25-32 16 32 Amoxicillin clavulanate0.06-4 0.5 4 Clindamycin

0.015->32

0.015 0.03 Erythromycin 0.06->32 1 2 Azithromycin 0.25->32 0.5 1Clarithromycin 0.06->32 0.125 0.25 Metronidazole 0.5-4 2 4 Prevotellabuccae-oris group [22]^(E) Gemifloxacin 0.5-8 2 2 Trovafloxacin 0.25-4 12 Penicillin G 0.06->32 8 >32 Amoxicillin clavulanate 0.125-2 0.25 1Erythromycin 0.5-8 1 2 Azithromycin 0.125-4 0.5 1 Clarithromycin 0.06-10.125 0.25 Clindamycin

0.015-0.125

0.015 0.03 Metronidazole 0.5-4 1 2 Prevotella heparinolytica [16]Gemifloxacin 0.25-0.5 0.5 0.5 Trovafloxacin 0.125-0.25 0.125 0.25Penicillin G 0.06-0.25 0.06 0.125 Amoxicillin clavulanate 0.06-0.250.125 0.25 Clindamycin

0.015

0.015

0.015 Erythromycin 0.25-0.5 0.25 0.25 Azithromycin 0.5-1 0.5 1Clarithromycin 0.06-0.125 0.125 0.125 Metronidazole 0.06-1 0.5 1Prevotella intermedia [11] Gemifloxacin 0.06-1 0.25 0.5 Trovafloxacin0.06-1 0.5 1 Penicillin G

0.015-16 0.03 4 Amoxicillin clavulanate 0.03-0.5 0.03 0.125 Clindamycin

0.015-0.03

0.015

0.015 Erythromcyin 0.03-0.5 0.06 0.25 Azithromycin 0.03-1 0.125 0.5Clarithromycin

0.015-0.125

0.015 0.125 Metronidazole 0.03-1 0.5 1 Prevotella melaninogenica [12]Gemifloxacin 0.125->8 1 8 Trovafloxacin 0.06-8 1 4 Penicillin G

0.015-2 0.25 2 Amoxicillin clavulanate 0.03-16 2 4 Clindamycin

0.015-32

0.015 0.5 Erythromycin 0.06-32 1 8 Azithromycin 0.125->32 0.25 32Clarithromycin 0.06-4 0.125 1 Metronidazole 0.125-4 0.5 1 Prevotelladenticola/ loeschii group [6] Gemifloxacin 0.25-8 0.5 Trovofloxacin0.06-4 1 Penicillin G

0.015-32 4 Amoxicillin clavulanate 0.03-0.5 0.06 Clindamycin

0.015-0.25

0.015 Erythromycin 0.125-16 0.25 Azithromycin 0.06-16 0.5 Clarithromycin0.03-2 0.06 Metronidazole 0.5-1 1 Veillonella spp. [24] Gemifloxacin0.03->8 1 8 Trovafloxacin 0.125->8 0.25 >8 Penicillin G

0.015-8 1 4 Amoxicillin clavulanate

0.015->4 0.5 2 Clindamycin 0.03->32 0.06 2 Erythromycin 1->32 16 >32Azithromycin 0.125->32 4 >32 Clarithromycin 1->32 16 >32 Metronidazole0.25-2 1 2

[0022] The invention provides a method for modulating metabolism ofanaerobic pathogenic bacteria. Skilled artisans can readily chooseanaerobic pathogenic bacteria or patients infected with or suspected tobe infected with these organisms to practice the methods of theinvention. Alternatively, the bacteria useful in the methods of theinvention may be those described herein.

[0023] The contacting step in any of the methods of the invention may beperformed in many ways that will be readily apparent to the skilledartisan. However, it is preferred that the contacting step is aprovision of a composition comprising a gemifloxacin compound to a humanpatient in need of such composition or directly to bacteria in culturemedium or buffer.

[0024] For example, when contacting a human patient or contacting saidbacteria in a human patient or in vitro, the compositions comprising aquinolone, particularly a gemifloxacin compound, preferablypharmaceutical compositions may be administered in any effective,convenient manner including, for instance, administration by topical,oral, anal, vaginal, intravenous, intraperitoneal, intramuscular,subcutaneous, intranasal or intradermal routes among others.

[0025] It is also preferred that these compositions be employed incombination with a non-sterile or sterile carrier or carriers for usewith cells, tissues or organisms, such as a pharmaceutical carriersuitable for administration to a subject. Such compositions comprise,for instance, a media additive or a therapeutically effective amount ofa compound of the invention, a quinolone, preferably a gemifloxacincompound, and a pharmaceutically acceptable carrier or excipient. Suchcarriers may include, but are not limited to, saline, buffered saline,dextrose, water, glycerol, ethanol and combinations thereof. Theformulation should suit the mode of administration.

[0026] Quinolone compounds, particularly gemifloxacin compounds andcompostions of the methods of the invention may be employed alone or inconjunction with other compounds, such as bacterial efflux pump inhibtorcompounds or antibiotic compounds, particularly non-quinolone compounds,e.g., beta-lactam antibiotic compounds.

[0027] In therapy or as a prophylactic, the active agent of a method ofthe invention is preferably administered to an individual as aninjectable composition, for example as a sterile aqueous dispersion,preferably an isotonic one.

[0028] Alternatively, the gemifloxacin compounds or compositions in themethods of the invention may be formulated for topical application forexample in the form of ointments, creams, lotions, eye ointments, eyedrops, ear drops, mouthwash, impregnated dressings and sutures andaerosols, and may contain appropriate conventional additives, including,for example, preservatives, solvents to assist drug penetration, andemollients in ointments and creams. Such topical formulations may alsocontain compatible conventional carriers, for example cream or ointmentbases, and ethanol or oleyl alcohol for lotions. Such carriers mayconstitute from about 1% to about 98% by weight of the formulation; moreusually they will constitute up to about 80% by weight of theformulation.

[0029] For administration to mammals, and particularly humans, it isexpected that the antibacterially effective amount is a daily dosagelevel of the active agent from 0.001 mg/kg to 10 mg/kg, typically around0.1 mg/kg to 1 mg/kg, preferably about 1 mg/kg. A physician, in anyevent, will determine an actual dosage that is most suitable for anindividual and will vary with the age, weight and response of theparticular individual. The above dosages are exemplary of the averagecase. There can, of course, be individual instances where higher orlower dosage ranges are merited, and such are within the scope of thisinvention. It is preferred that the dosage is selected to modulatemetabolism of the bacteria in such a way as to inhibit or stop growth ofsaid bacteria or by killing said bacteria. The skilled artisan mayidentify this amount as provided herein as well as using other methodsknown in the art, e.g. by the application MIC tests.

[0030] A further embodiment of the invention provides for the contactingstep of the methods to further comprise contacting an in-dwelling devicein a patient. In-dwelling devices include, but are not limited to,surgical implants, prosthetic devices and catheters, i.e., devices thatare introduced to the body of an individual and remain in position foran extended time. Such devices include, for example, artificial joints,heart valves, pacemakers, vascular grafts, vascular catheters,cerebrospinal fluid shunts, urinary catheters, and continuous ambulatoryperitoneal dialysis (CAPD) catheters.

[0031] A quinolone, particularly a gemifloxacin compound or compositionof the invention may be administered by injection to achieve a systemiceffect against relevant bacteria, preferably a anaerobic pathogenicbacteria, shortly before insertion of an in-dwelling device. Treatmentmay be continued after surgery during the in-body time of the device. Inaddition, the composition could also be used to broaden perioperativecover for any surgical technique to prevent bacterial wound infectionscaused by or related to anaerobic pathogenic bacteria.

[0032] In addition to the therapy described above, a gemifloxacincompound or composition used in the methods of this invention may beused generally as a wound treatment agent to prevent adhesion ofbacteria to matrix proteins, particularly anaerobic pathogenic bacteria,exposed in wound tissue and for prophylactic use in dental treatment asan alternative to, or in conjunction with, antibiotic prophylaxis.

[0033] Alternatively, a quinolone, particularly a gemifloxacin compoundor composition of the invention may be used to bathe an indwellingdevice immediately before insertion. The active agent will preferably bepresent at a concentration of 1 μg/ml to 10 μg/ml for bathing of woundsor indwelling devices.

[0034] Also provided by the invention is a method of treating orpreventing a bacterial infection by anaerobic pathogenic bacteriacomprising the step of administering an antibacterially effective amountof a composition comprising a quinolone, particularly a gemifloxacincompound to a mammal, preferably a human, suspected of having or beingat risk of having an infection with anaerobic pathogenic bacteria.

[0035] While a preferred object of the invention provides a methodwherein said anaerobic pathogenic bacteria is selected from the groupconsisting of: selected from the group consisting of: a member of thegenus Peptostreptococci, a Peptostreptococci asaccharolyticus, aPeptostreptococci magnus, a Peptostreptococci micros, aPeptostreptococci prevotii, a member of the genus Porphyromonas, aPorphyromonas asaccharolytica, a Porphyromonas canoris, a Porphyromonasgingivalis, a Porphyromonas macaccae, a member of the genus Actinomyces,an Actinomyces israelii, an Actinomyces odontolyticus, a member of thegenus Clostridium, a Clostridium innocuum, a Clostridiumclostridioforme, a Clostridium difficile, a member of the genusAnaerobiospirillum, a member of the genus Bacteroides, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a member of the genus Prevotella, a Prevotella intermedia, aPrevotella heparinolytica, a Prevotella oris-buccae, a Prevotella bivia,a Prevotella melaninogenica, a member of the genus Fusobacterium, aFusobacterium naviforme, a Fusobacterium necrophorum, a Fusobacteriumvarium, a Fusobacterium ulcerans, a Fusobacterium russii, a member ofthe genus Bilophila, a Bilophila wadsworthia.

[0036] Other anaerobic pathogenic bacteria may also be included in themethods. The skilled artisan may identify these organisms as providedherein as well as using other methods known in the art, e.g. MIC tests.

[0037] Preferred embodiments of the invention include, among otherthings, methods wherein said composition comprises gemifloxacin, or apharmaceutically acceptable derivative thereof.

EXAMPLES

[0038] The present invention is further described by the followingexamples. The examples are provided solely to illustrate the inventionby reference to specific embodiments. This exemplification's, whileillustrating certain specific aspects of the invention, do not portraythe limitations or circumscribe the scope of the disclosed invention.

[0039] All examples were carried out using standard techniques, whichare well known and routine to those of skill in the art, except whereotherwise described in detail.

[0040] All parts or amounts set out in the following examples are byweight, unless otherwise specified.

Example 1 Bacterial Strains

[0041] The strains were previously isolated from human clinicalspecimens from a variety of sources, and were identified by standardcriteria (Alexander, et al., J. Clin. Microbiol., 35:406-411, 1997;Citron, et al., Clin. Infect. Dis., 23 (Suppl. 1): 78-82, 1996;Holdeman, et al., Anaerobic Laboratory Manual, 4th Edition, 1977;Summanen, et al., Wadsworth Anaerobic Bacteriology Manual, 5th Edition,1993). Almost all these isolates were different from those strains usedin our prior study (Goldstein, et al., Antimicrob. Agents Chemother.,Submitted] when we used the same genus and species. Bacteroides fragilisATCC 25285, and Bacteroides thetaiotaomicron ATCC 29741 were testedsimultaneously as control strains. The numbers and species of isolatestested are given in Table 1.

Example 1 Compounds

[0042] Frozen cultures were transferred at least twice on Brucella agarsupplemented with hemin, vitamin K₁, and 5% sheep blood to ensure purityand good growth. Susceptibility testing was performed according toNational Committee for Clinical Laboratory Standards (NCCLS) standards(National Committee for Clinical Laboratory Standards, 4th Edition,1997). Brucella agar supplemented with hemin, vitamin K₁, and 5% lakedsheep blood was the basal medium used. For Bilophila wadsworthia, theagar was also supplemented with pyruvate. Antimicrobial agents werereconstituted according to the manufacturers' instructions. Serialtwofold dilutions of antimicrobial agents were prepared on the day ofthe test and added to the media in varying concentrations (

g/ml).

[0043] The agar plates were inoculated with a Steers replicator (CraftMachine Inc., Chester, Pa.). The inoculum used was 10⁵ CFU per spot.Control plates without antimicrobial agents were inoculated before andafter each set of drug-containing plates. The MIC was defined as thelowest concentration of an agent that yielded no growth, or a markedchange in the appearance of growth as compared to the growth controlplate.

[0044] Each reference cited herein is hereby incorporated by referencein its entirety. Moreover, each patent application to which thisapplication claims priority is hereby incorporated by reference in itsentirety.

What is claimed is:
 1. A method for modulating metabolism of anaerobicpathogenic bacteria comprising the step of contacting anaerobicpathogenic bacteria with an antibacterially effective amount of acomposition comprising a gemifloxacin compound, or antibacteriallyeffective derivatives thereof.
 2. The method of claim 1 wherein saidanaerobic pathogenic bacteria is selected from the group consisting of:a member of the genus Peptostreptococci, a Peptostreptococciasaccharolyticus, a Peptostreptococci magnus, a Peptostreptococcimicros, a Peptostreptococci prevotii, a Porphyromonas asaccharolytica, aPorphyromonas canoris, a Porphyromonas gingivalis, a Porphyromonasmacaccae, a member of the genus Actinomyces, an Actinomyces israelii, anActinomyces odontolyticus, a member of the genus Clostridium, aClostridium innocuum, a Clostridium clostridioforme, a Clostridiumdifficile, a member of the genus Anaerobiospirillum, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a Prevotella intermedia, a Prevotella heparinolytica, aPrevotella oris-buccae, a Prevotella bivia, a Prevotella melaninogenica,a member of the genus Fusobacterium, a Fusobacterium naviforme, aFusobacterium necrophorum, a Fusobacterium varium, a Fusobacteriumulcerans, a Fusobacterium russii, a member of the genus Bilophila, aBilophila wadsworthia.
 3. A method of treating or preventing a bacterialinfection by anaerobic pathogenic bacteria comprising the step ofadministering an antibacterially effective amount of a compositioncomprising a gemifloxacin compound to a mammal suspected of having orbeing at risk of having an infection with anaerobic pathogenic bacteria.4. The method of claim 3 wherein said anaerobic pathogenic bacteria isselected from the group consisting of: a member of the genusPeptostreptococci, a Peptostreptococci asaccharolyticus, aPeptostreptococci magnus, a Peptostreptococci micros, aPeptostreptococci prevotii, a Porphyromonas asaccharolytica, aPorphyromonas canoris, a Porphyromonas gingivalis, a Porphyromonasmacaccae, a member of the genus Actinomyces, an Actinomyces israelii, anActinomyces odontolyticus, a member of the genus Clostridium, aClostridium innocuum, a Clostridium clostridioforme, a Clostridiumdifficile, a member of the genus Anaerobiospirillum, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a Prevotella intermedia, a Prevotella heparinolytica, aPrevotella oris-buccae, a Prevotella bivia, a Prevotella melaninogenica,a member of the genus Fusobacterium, a Fusobacterium naviforme, aFusobacterium necrophorum, a Fusobacterium varium, a Fusobacteriumulcerans, a Fusobacterium russii, a member of the genus Bilophila, aBilophila wadsworthia.
 5. The method of claim 1 wherein said modulatingmetabolism is inhibiting growth of said bacteria.
 6. The method of claim1 wherein said modulating metabolism is killing said bacteria.
 7. Themethod of claim 1 wherein said contacting said bacteria comprises thefurther step of introducing said composition into a mammal.
 8. Themethod of claim 3 wherein said mammal is a human.
 9. The method of claim7 wherein said mammal is a human.
 10. The method of claim 1 wherein saidbacteria is selected from the group consisting of: a member of the genusPeptostreptococci, a Peptostreptococci asaccharolyticus, aPeptostreptococci magnus, a Peptostreptococci micros, aPeptostreptococci prevotii, a Porphyromonas asaccharolytica, aPorphyromonas canoris, a Porphyromonas gingivalis, a Porphyromonasmacaccae, a member of the genus Actinomyces, an Actinomyces israelii, anActinomyces odontolyticus, a member of the genus Clostridium, aClostridium innocuum, a Clostridium clostridioforme, a Clostridiumdifficile, a member of the genus Anaerobiospirillum, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a Prevotella intermedia, a Prevotella heparinolytica, aPrevotella oris-buccae, a Prevotella bivia, a Prevotella melaninogenica,a member of the genus Fusobacterium, a Fusobacterium naviforme, aFusobacterium necrophorum, a Fusobacterium varium, a Fusobacteriumulcerans, a Fusobacterium russii, a member of the genus Bilophila, aBilophila wadsworthia.
 11. The method of claim 1 wherein said bacteriais selected from the group consisting of: a member of the genusPeptostreptococci, a Peptostreptococci asaccharolyticus, aPeptostreptococci magnus, a Peptostreptococci micros, aPeptostreptococci prevotii, a Porphyromonas asaccharolytica, aPorphyromonas canoris, a Porphyromonas gingivalis, a Porphyromonasmacaccae, a member of the genus Actinomyces, an Actinomyces israelii, anActinomyces odontolyticus, a member of the genus Clostridium, aClostridium innocuum, a Clostridium clostridioforme, a Clostridiumdifficile, a member of the genus Anaerobiospirillum, a Bacteroidestectum, a Bacteroides ureolyticus, a Bacteroides gracilis (Campylobactergracilis), a Prevotella intermedia, a Prevotella heparinolytica, aPrevotella oris-buccae, a Prevotella bivia, a Prevotella melaninogenica,a member of the genus Fusobacterium, a Fusobacterium naviforme, aFusobacterium necrophorum, a Fusobacterium varium, a Fusobacteriumulcerans, a Fusobacterium russii, a member of the genus Bilophila, aBilophila wadsworthia.